Blasticidin S inhibits translation by trapping deformed tRNA on the ribosome

Svidritskiy, Egor; Ling, Clarence; Ermolenko, Dmitri N.; Коростелев, А.A.

doi:10.1073/pnas.1304922110

Cited by 91 publications

(119 citation statements)

References 49 publications

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“…S6). While these rearrangements are reminiscent of those observed for P-tRNA in the presence of the antibiotic blasticidin S (Svidritskiy et al, 2013) and for deacylated tRNA bound at the E site (Schmeing et al, 2003; Selmer et al, 2006) (Fig. S6), in these latter cases the conformational changes are local and do not disturb the overall conformation of the acceptor arm.…”

Section: Resultsmentioning

confidence: 64%

Distinct tRNA Accommodation Intermediates Observed on the Ribosome with the Antibiotics Hygromycin A and A201A

Starosta²,

et al. 2015

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SUMMARY The increase in multi-drug resistant bacteria is limiting the effectiveness of currently approved antibiotics, leading to a renewed interest in antibiotics with distinct chemical scaffolds. We have solved the structures of the Thermus thermophilus 70S ribosome with A-, P- and E-site tRNAs bound, and in complex with either the aminocyclitol-containing antibiotic hygromycin A (HygA) or the nucleoside antibiotic A201A. Both antibiotics bind at the peptidyl transferase center and sterically occlude the CCA-end of the A-tRNA from entering the A-site of the peptidyl transferase center. Single-molecule Förster resonance energy transfer (smFRET) experiments reveal that HygA and A201A specifically interfere with full accommodation of the A-tRNA, leading to the presence of tRNA accommodation intermediates, and thereby inhibiting peptide bond formation. Thus, our results provide not only insight into the mechanism of action of HygA and A201A, but also into the fundamental process of tRNA accommodation during protein synthesis.

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Section: Resultsmentioning

confidence: 64%

Distinct tRNA Accommodation Intermediates Observed on the Ribosome with the Antibiotics Hygromycin A and A201A

Starosta²,

et al. 2015

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“…The kinetics of mRNA translocation were followed by the fluorescence quenching of a fluorescein dye attached to the 3’ end of an mRNA as it moves within the ribosome[41, 42]. Pretranslocation complexes were assembled with fluorescein-labeled mRNA, deacylated tRNA Met , N -acetyl-Tyr-tRNA Tyr and 70S ribosomes.…”

Section: Resultsmentioning

confidence: 99%

EF-G Activation by Phosphate Analogs

Salsi

Farah

Ermolenko

2016

Journal of Molecular Biology

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EF-G is a universally conserved translational GTPase that promotes the translocation of tRNA and mRNA through the ribosome. EF-G binds to the ribosome in a GTP-bound form and subsequently catalyzes GTP hydrolysis. The contribution of the ribosome-stimulated GTP hydrolysis by EF-G to tRNA/mRNA translocation remains debated. Here, we show that while EF-G•GDP does not stably bind to the ribosome and induce translocation, EFG• GDP in complex with phosphate group analogues BeF3− and AlF4− promotes the translocation of tRNA and mRNA. Furthermore, the rates of mRNA translocation induced by EF-G in the presence of GTP and a non-hydrolysable analogue of GTP, GDP•BeF3−are similar. Our results are consistent with the model suggesting that GTP hydrolysis is not directly coupled to mRNA/tRNA translocation. Hence, GTP binding is required to induce the activated, translocation-competent conformation of EF-G while GTP hydrolysis triggers EF-G release from the ribosome.

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“…4a and Extended Data Fig. 7) 23 . Remarkably, chemically diverse inhibitors share a similar mode of binding within the pocket.…”

Section: The Peptidyl Transferase Centrementioning

confidence: 98%